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A study of proteolysis during Camembert cheese ripening using isoelectric focusing and two-dimensional electrophoresis

Published online by Cambridge University Press:  01 June 2009

Patrick Trieu-Cuot
Affiliation:
Laboratoire de Biochimie et Technologie Laitières, Institut National de la Recherche Agronomique CNRZ, 78350 Jouy-en-Josas, France
Jean-Claude Gripon
Affiliation:
Laboratoire de Biochimie et Technologie Laitières, Institut National de la Recherche Agronomique CNRZ, 78350 Jouy-en-Josas, France

Summaby

Isoelectric focusing and 2-dimensional electrophoresis were used to study the development of the pH 4·6-insoluble fraction during Camembert cheese ripening. Modifications of this fraction were due mainly to the action of 5 proteinases: rennet (chymosin + bovine pepsin), plasmin and Penicillium caseicolum aspartyl-and metalloproteinases. Rennet was inactive on β-casein, but acted very early on αs1-casein. However, rennet and P. caseicolum aspartyl-proteinase had a very similar action on the latter substrate, which prevented clear definition of the respective actions of these proteinases on αs1-casein after 7 d of ripening. Plasmin action on β-casein was important from 21 and 35 d of ripening at the surface and in the centre of the cheese respectively, suggesting an important influence of pH changes during maturation. The respective activities of the metallo-and aspartyl-proteinases of P. caseicolum were characterized and followed using β-casein degradation products as markers. The metallo-proteinase activity was detectable immediately after the development of the Penicillium (7 d), while that of the aspartyl-proteinase was observed 3 d later. Thereafter, the amount of β-casein degradation peptides resulting from the metalloproteinase decreased while that resulting from the aspartyl-proteinase increased, suggesting a more important role of the latter enzyme.

Type
Original Articles
Copyright
Copyright © Proprietors of Journal of Dairy Research 1982

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References

REFERENCES

Alais, C. 1974 [Biochemistry of enzymic coagulation of milk.] Chimia 28 597604Google Scholar
Berry, G. P. & Creamer, L. K. 1975 The association of bovine βcasein. The importance of the C-terminal region. Biochemistry 14 35423545CrossRefGoogle ScholarPubMed
Creamer, L. K. 1970 Protein breakdown in Gouda cheese. New Zealand Journal of Dairy Science and Technology 5 152154Google Scholar
Creamer, L. K. 1975 β-Casein degradation in Gouda and Cheddar cheese. Journal of Dairy Science 58 287292CrossRefGoogle Scholar
Desmazeaud, M. J., Gripon, J. C., D.Le Bars, Le Bars, & Beroere, J. L. 1976 [The role of microorganisms and enzymes in cheese maturation. 3. Effect of microorganisms.] Lait 56 379396CrossRefGoogle Scholar
Fox, P. F. & Walley, B. F. 1971 Influence of sodium chloride on the proteolysis of casein by rennet and by pepsin. Journal of Dairy Research 38 165170CrossRefGoogle Scholar
Green, M. L. & Foster, P. M. D. 1974 Comparison of the rates of proteolysis during ripening of Cheddar cheeses made with calf rennet and swine pepsin as coagulants. Journal of Dairy Research 41 269282CrossRefGoogle Scholar
Gripon, J. C, Desmazeaud, M. J., D., Le Bars & Beroere, J. L. 1975 [The role of microorganisms and enzymes in cheese maturation. 2. Effect of commercial rennet.] Lait 55 502516CrossRefGoogle Scholar
Gripon, J. C, Desmazeaud, M. J., D.Le Bars, Le Bars, & Beroere, J. L. 1977 Role of proteolytic enzymes of Streptococcus lactis, Penicillium roqueforti and Penicillium caseicolum during cheese ripening. Journal of Dairy Science 60 15321538CrossRefGoogle Scholar
Hofmann, C. J., Keenan, T. W. & Eigel, W. N. 1979 Association of plasminogen with bovine milk fat globule membrane. International Journal of Biochemistry 10 909917CrossRefGoogle ScholarPubMed
Ledford, R. A., O'Sullivan, A. C. & Nath, K. R. 1966 Residual casein fractions in ripened cheese determined by polyacrylamide-gel electrophoresis. Journal of Dairy Science 49 10981101CrossRefGoogle ScholarPubMed
Lenoir, J. 1963 [Note on the degradation of nitrogen compounds during ripening of Camembert cheese.] Lait 43 154165CrossRefGoogle Scholar
Lenoir, J. 1970 [Protease activity in soft cheeses of Camembert type.] Revue Laitière Française (no. 275) 231 233 235 237 239 241 243Google Scholar
Lenoir, J. 1979 Thesis, Université de CaenGoogle Scholar
Lenoir, J., Auberoer, B. & Gripon, J. C. 1979 [Characteristics of the proteolytic system of Penicillium caseicolum. 3. Characterization of an acid protease.] Lait 59 244268CrossRefGoogle Scholar
Mercier, J. C., Ribadeau-Dumas, B. & Grosclaude, F. 1973 Amino-acid composition and sequence of bovine κ-casein. Netherlands Milk and Dairy Journal 27 313322Google Scholar
Noomen, A. 1975 Proteolytic activity of milk protease in raw and pasteurized cow's milk. Netherlands Milk and Dairy Journal 29 153161Google Scholar
Noomen, A. 1978 a Activity of proteolytic enzymes in simulated soft cheeses (Meshanger type). 1. Activity of milk protease. Netherlands Milk and Dairy Journal 32 2648Google Scholar
Noomen, A. 1978 b Activity of proteolytic enzymes in simulated soft cheeses (Meshanger type). 2. Activity of calf rennet. Netherlands Milk and Dairy Journal 32 4968Google Scholar
O'Keeffe, R. B., Fox, P. F. & Daly, C. 1976 Contribution of rennet and starter proteases to proteolysis in Cheddar cheese. Journal of Dairy Research 43 97107CrossRefGoogle Scholar
Reimerdes, E. H. & Herlitz, E. 1979 The formation of γ-caseins during cooling of raw milk. Journal of Dairy Research 46 219221CrossRefGoogle ScholarPubMed
Reiter, B., Sorokin, Y., Pickering, A. & Hall, A. J. 1969 Hydrolysis of fat and protein in small cheeses made under aseptic conditions. Journal of Dairy Research 36 6576CrossRefGoogle Scholar
B.Ribadeau Dumas, Ribadeau Dumas,, Brignon, G.Grosclaude, F. & Mercier, J. C. 1972 [Primary structure of bovine β-casein. Complete sequence.] European Journal of Biochemistry 25 505514Google Scholar
Righetti, P. G., Muneroni, P., Todesco, R. & Carini, S. 1980 Fingerprinting of casein digests by isoelectric focusing and SDS electrophoresis. Electrophoresis 1 3742CrossRefGoogle Scholar
Snoeren, T. H. M. & M.Van Riel, J. A. Van Riel, J. A. 1979 Milk proteinase, its isolation and action on αs2- and β-casein. Milchwissenschaft 34 528531Google Scholar
Trieu-Cuot, P. 1981 Thesis. Université Paris-Sud – Centre d'OrsayGoogle Scholar
Trieu-Cuot, P., Archieri-Haze, M. J. & Gripon, J. C. 1982 a Effect of aspartyl proteinases of Penicillium caseicolum and Penicillium roqueforti on caseins. Journal of Dairy Research 49 487500CrossRefGoogle Scholar
Trieu-Cuot, P., Archieri-Haze, M. J. & Gripon, J. C. 1982 b [Comparison of hydrolysis of αs1- and β-casein by metalloproteinase of Penicillium caseicolum and Penicillium roqueforti.] Lait 62 (in press)Google Scholar
Trieu-Cuot, P. & Gripon, J. C. 1981 Electrofocusing and two-dimensional electrophoresis of bovine caseins. Journal of Dairy Research 48 303310CrossRefGoogle Scholar
Veisseyre, R. 1974 Techniques laitières. Paris: La Maison RustiqueGoogle Scholar
Visser, F. M. W. & A.De Groot-Mostert, A. E. De Groot-Mostert, A. E. 1977 Contribution of enzymes from rennet, starter bacteria and milk to proteolysis and flavour development in Gouda cheese. 4. Protein breakdown: a gel electrophoretical study. Netherlands Milk and Dairy Journal 31 247264Google Scholar